Differences in pea (Pisum sativum L.) and chickpea (Cicer arietinum L.) microbial compatibility and/ or their associated farming practices may influence root fungi of the following crop and affect the yield. The main objective of this research was to explain the difference in durum wheat (Triticum turgidum L.) yield the year after pea and chickpea crops through changes in the functional diversity of wheat root fungi. The effect of fungicides used on chickpea on the root fungi of a following durum wheat crop was studied using plate culture and pyrosequencing. Pyrosequencing detected more Fusarium spp. in the roots of durum wheat after fungicide-treated chickpea than in non-fungicide treated chickpea. Plate culture revealed that the functional groups of fungi responded differently to fungicide use in the field but the effect on total community was non-significant. Highly virulent pathogens were not affected, but antagonists were suppressed. More fungal antagonists were detected after the chickpea CDC Luna than CDC Vanguard. Fungal species responded differently to the use of fungicides in vitro, but the aggregate inhibition effect on antagonists and highly virulent pathogens was similar. The effect of chickpea vs. pea previous crop and different chickpea termination times on root fungi of a following durum wheat crop was studied. The abundance of Fusarium spp. increased after cultivation of both cultivars of chickpea as compared to pea according to pyrosequencing and was negatively correlated with durum yield. Plate culture analysis revealed that fungal antagonists were more prevalent after pea than both cultivars of chickpea and chickpea CDC Vanguard increased the abundance of highly virulent pathogens. The abundance of highly virulent pathogens in durum wheat roots was negatively correlated to durum yield. Early termination of chickpea did not change the community of culturable fungi in the roots of a following durum crop. It is noteworthy that Fusarium redolens was identified for the first time in Saskatchewan and its pathogenicity was confirmed on durum wheat, pea and chickpea. The classical method of root disease diagnostics in cereals is based on the examination of the subcrown internode. I evaluated the method by comparing the fungal communities associated with different subterranean organs of durum wheat. The fungal community of the subcrown internode was different from that of roots and crown, suggesting cautious use of this method.

Characteristics of quantitative resistance in pea (Pisum sativum L.) to Erysiphe pisi DC, the pathogen causing powdery mildew, were investigated. Cultivars and seedlines of pea expressing quantitative resistance to E. pisi were identified and evaluated, by measuring the amounts of pathogen present on plant surfaces in field and glasshouse experiments. Disease severity on cv. Quantum was intermediate when compared with that on cv. Bolero (susceptible) and cv. Resal (resistant) in a field experiment. In glasshouse experiments, two groups of cultivars, one with a high degree of resistance and the other with nil to low degrees of resistance to E. pisi, were identified. This indicated either that a different mechanism of resistance applied in the two groups, or that there has been no previous selection for intermediate resistance. Several other cultivars expressing quantitative resistance were identified in a field experiment. Quantitative resistance in Quantum did not affect germination of E. pisi conidia, but reduced infection efficiency of conidia on this cultivar compared with cv. Pania (susceptible). Other epidemiological characteristics of quantitative resistance expression in Quantum relative to Pania were a 33% reduction in total conidium production and a 16% increase in time to maximum daily conidium production, both expressed on a colony area basis. In Bolero, the total conidium production was reduced relative to Pania, but the time to maximum spore production on a colony area basis was shorter. There were no differences between the cultivars in pathogen colony size or numbers of haustoria produced by the pathogen. Electron microscope studies suggested that haustoria in Quantum plants were smaller and less lobed than those in Pania plants and the surface area to volume ratios of the lobes and haustorial bodies were larger in Pania than in Quantum. The progress in time and spread in space of E. pisi was measured in field plots of cultivars Quantum, Pania and Bolero as disease severity (proportion of leaf area infected). Division of leaves (nodes) into three different age groups (young, medium, old) was necessary because of large variability in disease severity within plants. Disease severity on leaves at young nodes was less than 4% until the final assessment at 35 days after inoculation (dai). Exponential disease progress curves were fitted for leaves at medium nodes. Mean disease severity on medium nodes 12 dai was greatest (P<0.001) on Bolero and Pania (9.3 and 6.8% of leaf area infected respectively), and least on Quantum (1.6%). The mean disease relative growth rate was greatest (P<0.001) for Quantum, but was delayed compared to Pania and Bolero. Gompertz growth curves were fitted to disease progress data for leaves at old nodes. The asymptote was 78.2% of leaf area infected on Quantum, significantly lower (P<0.001) than on Bolero or Pania, which reached 100%. The point of inflection on Quantum occurred 22.8 dai, later (P<0.001) than on Pania (18.8 dai) and Bolero (18.3 dai), and the mean disease severity at the point of inflection was 28.8% for Quantum, less (P<0.00l) than on Pania (38.9%) or Bolero (38.5%). The average daily rates of increase in disease severity did not differ between the cultivars. Disease progress on Quantum was delayed compared with Pania and Bolero. Disease gradients from inoculum foci to 12 m were detected at early stages of the epidemic but the effects of background inoculum and the rate of disease progress were greater than the focus effect. Gradients flattened with time as the disease epidemic intensified, which was evident from the large isopathic rates (between 2.2 and 4.0 m d⁻¹) Some epidemiological variables expressed in controlled environments (low infection efficiency, low maximum daily spore production and long time to maximum spore production) that characterised quantitative resistance in Quantum were correlated with disease progress and spread in the field. These findings could be utilised in pea breeding programmes to identify parent lines from which quantitatively resistant progeny could be selected.

[ES] Las plantas monocárpicas se definen como aquellas que florecen, producen semillas y mueren después de un solo ciclo reproductivo. Muchos cultivos de importancia agroeconómica siguen estrategia reproductiva. En estas plantas, después de producir un cierto número de semillas, el meristemo apical del tallo (SAM) cesa su actividad, siendo esta la antesala de su senescencia y muerte. Este fenómeno, estudiado en diferentes especies de plantas monocárpicas, se conoce como Parada Proliferativa (PA). Se encuentra influenciado por múltiples factores, que incluyen la influencia del desarrollo de frutos y semillas, así como las condiciones ambientales de luz, humedad, etc. Todos estos factores son finalmente integrados a nivel genético en la planta. En este contexto, en Aabidopsis thaliana se ha descrito una ruta dependiente de la edad encargada de la modulación del PA, la ruta FUL-AP2. Se ha demostrado que el miR172 y FRUITFUL (FUL) incrementan su expresión con la edad de la planta y regulan negativamente APETALA2 (AP2), que es responsable de mantener la actividad meristemática a través de la acción de WUSCHEL (WUS). En este sentido, se ha sugerido que otros miembros de la subfamilia euAP2, TOE1, TOE2, TOE3, SMZ y SNZ, conocidos colectivamente como AP2-like genes, también juegan un papel crucial en la modulación de la PA. No obstante, estos han sido definidos principalmente como reguladores de la transición floral, y su implicación en la modulación del PA no está bien establecida. Por otro lado, Pisum sativum ha sido, históricamente, una especie ampliamente estudiada a nivel fisiológico en lo que concierne a PA. Sin embargo, hoy aún existen numerosas cuestiones, ambigüedades y discrepancias acerca de la regulación del PA en esta especie. En esta tesis doctoral, pretendemos profundizar en la caracterización de diversos aspectos del PA tanto en Arabidopsis thaliana como en Pisum sativum, con intención de determinar el grado de conservación que existe en este proceso entre estas dos especies monocárpicas. En el Capítulo 1, hemos examinado el papel de todos los miembros de la subfamilia euAP2 en la modulación de la PA en Arabidopsis thaliana, así como su potencial para estrategias biotecnológicas dirigidas a la modulación de la PA. Nuestros resultados sugieren que, a excepción de SMZ, todos juegan un papel crítico en este proceso, siendo inductores de la actividad meristemática. Además, AP2 y SNZ han demostrado tener el potencial para ser usados en estrategias biotecnológicas dirigidas a aumentar la producción de frutos en plantas monocárpicas. En el capítulo 2 se han revisitado los estudios fisiológicos que históricamente han tenido como objetivo determinar el papel de los frutos en desarrollo en la inducción del PA en Pisum sativum. Nuestros resultados sugieren que el desarrollo de semillas determina el momento del PA: cuando se ha producido una determinada biomasa de semillas, la SAM entra en un estado latente. Además, a nivel transcriptómico, Arabidopsis thaliana y Pisum sativum exhiben un comportamiento similar en cuanto a la influencia de las semillas en la actividad meristemática y el PA. Finalmente, en el Capítulo 3, hemos generado herramientas para caracterizar los genes de la subfamilia euAP2 en la modulación de PA en Pisum sativum. Además, sentamos las bases para el estudio de nuevos moduladores de la ruta genética, como Flowering Locus T (FT), postulándolo así como posible florígeno y anti-florígeno de las plantas. / [CA] Les plantes monocàrpiques es definixen com aquelles que florixen, produïxen llavors i moren després d'un sol cicle reproductiu. Molts cultius d'importància agroeconómica seguixen estratègia reproductiva. En estes plantes, després de produir un cert nombre de llavors, el meristemo apical de la tija (SAM) cessa la seua activitat, sent esta l'avantsala de la seua senescència i mort. Este fenomen, estudiat en diferents espècies de plantes monocàrpiques, es coneix com a Parada Proliferativa (PA). Es troba influenciat per múltiples factors, que inclouen la influència del desenvolupament de fruits i llavors, així com les condicions ambientals de llum, humitat, etc. Tots estos factors són finalment integrats a nivell genètic en la planta. En este context, en Aabidopsis thaliana s'ha descrit una ruta dependent de l'edat encarregada de la modulació del PA, la ruta FUL-AP2. S'ha demostrat que el miR172 i FRUITFUL (FUL) incrementen la seua expressió amb l'edat de la planta i regulen negativament APETALA2 (AP2), que és responsable de mantindre l'activitat meristemàtica a través de l'acció de WUSCHEL (WUS). En este sentit, s'ha suggerit que altres membres de la subfamília euAP2, TOE1, TOE2, TOE3, SMZ i SNZ, coneguts col·lectivament com AP2-like gens, també juguen un paper crucial en la modulació de la PA. No obstant això, estos han sigut definits principalment com a reguladors de la transició floral, i la seua implicació en la modulació del PA no està ben establida. D'altra banda, Pisum sativum ha sigut, històricament, una espècie àmpliament estudiada a nivell fisiològic en el que concernix PA. No obstant això, hui encara existixen nombroses qüestions, ambigüitats i discrepàncies sobre la regulació del PA en esta espècie. En esta tesi doctoral, pretenem aprofundir en la caracterització de diversos aspectes del PA tant en Arabidopsis thaliana com en Pisum sativum, amb intenció de determinar el grau de conservació que existix en este procés entre estes dos espècies monocàrpiques. En el Capítol 1, hem examinat el paper de tots els membres de la subfamília euAP2 en la modulació de la PA en Arabidopsis thaliana, així com el seu potencial per a estratègies biotecnològiques dirigides a la modulació de la PA. Els nostres resultats suggerixen que, a excepció de SMZ, tots juguen un paper crític en este procés, sent inductors de l'activitat meristemàtica. A més, AP2 i SNZ han demostrat tindre el potencial per a ser usats en estratègies biotecnològiques dirigides a augmentar la producció de fruits en plantes monocàrpiques. En el capítol 2 s'han revisitat els estudis fisiològics que històricament han tingut com a objectiu determinar el paper dels fruits en desenvolupament en la inducció del PA en Pisum sativum. Els nostres resultats suggerixen que el desenvolupament de llavors determina el moment del PA: quan s'ha produït una determinada biomassa de llavors, la SAM entra en un estat latent. A més, a nivell transcriptómico, Arabidopsis thaliana i Pisum sativum exhibixen un comportament similar quant a la influència de les llavors en l'activitat meristemàtica i el PA. Finalment, en el Capítol 3, hem generat ferramentes per a caracteritzar els gens de la subfamília euAP2 en la modulació de PA en Pisum sativum. A més, establim les bases per a l'estudi de nous moduladors de la ruta genètica, com Flowering Locus T (FT), postulant-lo així com possible florígeno i anti-florígeno de les plantes. / [EN] Monocarpic plants are defined as those that bloom, produce seeds, and die after a single reproductive cycle. Many economically important crops belong to this reproductive strategy. In these plants, after producing a certain number of seeds, the shoot apical meristem (SAM) ceases its activity, heralding the onset of senescence and plant death. This phenomenon, studied in various monocarpic plant species, is known as Proliferative Arrest (PA), and is found to be influenced by multiple factors, involving the influence of developing fruits and seeds, as well as environmental conditions like temperature, light, and humidity. All these factors are ultimately integrated at the genetic level within the plant. In this context, an age-dependent pathway that controls SAM activity and modulates PA, known as the FUL-AP2 pathway, has been described in Arabidopsis thaliana. It has been demonstrated that the microRNA miR172 and FRUITFUL (FUL) increase with the plant's age and negatively regulate APETALA2 (AP2), which is responsible for maintaining meristematic activity through the action of WUSCHEL (WUS). In this sense, it has been suggested that other members of the euAP2 subfamily, TARGET OF EAT1 (TOE1), TOE2, TOE3, SCHLAFMÜTZE (SMZ), and SCHNARCHZAPFEN (SNZ), collectively known as AP2-like genes, also play a crucial role in modulating PA. However, they have mainly been defined as regulators of the floral transition, and their involvement in PA modulation is not well established. On the other hand, historically, Pisum sativum has been a species widely studied in relation to PA. However, this research often treated PA and senescence as the same process, and uncertainties and ambiguities persist, with discrepancies among different research groups that have treated this topic. In this doctoral thesis, we aimed to delve into the characterization of various aspects of PA in both Arabidopsis thaliana and Pisum sativum and determine the degree of conservation of this process in these two monocarpic species: In Chapter 1, we examined the role of all euAP2 subfamily members in modulating PA in Arabidopsis thaliana and explored their potential for biotechnological strategies aimed at PA modulation. Our findings suggest that, except for SMZ, all members of the euAP2 subfamily play a critical role in this process as inducers of meristematic activity. Furthermore, AP2 and SNZ have shown the potential to be considered prime candidates for use in biotechnological strategies to increase fruit production in monocarpic plants. Chapter 2 revisited the physiological studies that have historically aimed to determine the role of developing fruits in inducing PA in Pisum sativum. Our results suggest that developing seeds determine the timing of PA: when a certain seed biomass has been produced, the SAM enters a dormant state. Additionally, at the transcriptomic level, Arabidopsis thaliana and Pisum sativum exhibit similar behaviour regarding the influence of seeds on meristematic activity, suggesting that PA could be a conserved process among monocarpic plants. In Chapter 3, we generated tools for characterizing euAP2 subfamily genes in PA modulation in Pisum sativum. Furthermore, we laid the groundwork for the study of new modulators of the genetic pathway, such as Flowering Locus T (FT), suggesting its potential role of florigen and anti-florigen of the plant. / Burillo Richart, E. (2024). Characterization of Proliferative Arrest (PA) Process in Arabidopsis thaliana and Pisum sativum [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/207109